Flow cytometric assessment of ethylene glycol monoethyl ether on spermatogenesis in rats

The effects of ethylene glycol monoethyl ether (EGEE) on testicular cell populations in rats were investigated by a flow cytometric method. Rats were administered by gavage with EGEE at the various doses of 0 (saline alone), 100, 200, 400, and 800 mg/kg body weight/day for 4 weeks. The treatment of EGEE caused decreases in the weight of testis and epididymis and in the number of testicular cells. Histopathologically, exfoliation of germ cells into the tubular lumen was observed at the doses of above 200 mg/kg. The treatment of EGEE at the dose of 400 mg/kg caused moderate testicular degeneration. A significant depletion of haploid cells and a disproportionate ratio of diploid and tetraploid cells were observed as determined by flow cytometric analysis. These results indicate that the toxic effect of EGEE on the male reproductive system may be strongly associated with the disproportion of testicular germ cells.     

Effects of dietary electrolyte balance on the chemistry of blood and urine in lactating sows and sow litter performance

One hundred fifty-three sows (average parity of 2.2) were used to determine the effects of dietary electrolyte balance (calculated as mEq/kg of diet for Na + K - Cl) on sows and their litters during lactation. The sows were fed corn-soybean meal-based diets (1.0% lysine, 1.0% valine, 0.95% Ca, and 0.80% P; as-fed basis) starting on d 109 of gestation and throughout the 21-d lactation experiment. Dietary electrolyte balance (dEB) was 0, 100, 200, 350, and 500 mEq/kg (as-fed basis), well above and below the dEB of 185 mEq/kg found in a simple corn-soybean meal-based lactation diet. To achieve the desired dEB, diets had the following: 1) 1.8% HCl (6 N) and 1.06% CaCl2, 2) 1.0% CaCl2, 3) 0.04% NaHCO3, 4) 1.29% NaHCO3, and 5) 2.54% NaHCO3 (as-fed basis). Increasing dEB increased blood pH (linear and quadratic effects, P < 0.001), partial pressure of carbon dioxide (linear effect, P < 0.001), HCO3- concentration (linear and quadratic effects, P < 0.001), and blood base excess (linear and quadratic effects, P < 0.001). However, increased dEB resulted in lower blood concentrations of K (linear and quadratic effects, P < 0.04), Cl (linear and quadratic effects, P < 0.001), and ionized Ca (linear and quadratic effects, P < 0.001). Changing dEB did not affect ADFI; water usage, litter weight gain; sow weight change; sow backfat change; percentages of CP, lactose, and fat in the milk; percentage of sows returning to estrus; days to estrus; and number of pigs born alive in the subsequent litter (P = 0.06). However, piglet survivability to d 10 and overall was greatest with the lower dEB treatments (linear effect, P < 0.05). The pH (linear and quadratic effects, P < 0.001) and colony forming units of total bacteria (linear effect, P < 0.03) in the urine increased as dEB of the diet was increased. In conclusion, dEB had pronounced effects on the physiological status of sows and decreasing dEB below that in a simple corn-soybean meal-based diet decreased bacterial counts in the urine and increased piglet survivability. However, milk composition, sow and litter weights at weaning, and subsequent rebreeding performance of the sows were not affected by dEB.     

Intestinal immunomodulation by vitamin A deficiency and lactobacillus-based probiotic in Eimeria acervulina-infected broiler chickens

In a 2 X 2 factorial study, a broiler starter ration was amended for vitamin A (control, C; deficient, A) and probiotic status (-, P) to investigate their modulatory effects onthe host immune system. Birds were inoculated orally with Eimeria acervulina (EA) oocysts, and disease susceptibility was evaluated by assessment of fecal oocyst shedding. Humoral and local cellular mediated immunity were assessed by evaluation of antibody and cytokine (interferon-gamma [IFN-gamma] and interleukin-2 [IL-2]) levels in sera and intestinal secretions on a 3-day interval after inoculation. Fecal oocyst shedding was highest (P < 0.05) in A- birds, followed by AP, C-, and CP birds. Feeding the probiotic reduced shed oocysts by 20% in A fed birds and by 26% in C fed birds. Intestinal IFN-gamma was relatively constant in all treatment groups except for A-, where it declined steadily and was lower (P < 0.05) from day 6 on. Serum IFN-gamma levels fluctuated within each treatment and over time were not revealing. Intestinal IL-2 was highest in CP birds at 3 and 9 days postinfection (DPI) and lowest in A- birds at 3, 9, and 12 DPI (P < 0.05); no difference between treatments was found at 6 DPI (P > 0.05). Eimeria-specific intestinal antibody (Ab) level was constant (P > 0.05) in C- birds but increased with time (P < 0.05) in A-, AP, and CP birds. Serum Ab levels were also constant in A- and CP birds but increased (P < 0.05) in C- and AP birds after 6 DPI. The data demonstrate for the first time a probiotic-enhanced immunity in vitamin A deficient birds. This study is also the first to demonstrate the probiotic effect on local cell-mediated immunity of chickens, best manifested by apparent lower intestinal invasion and development by EA, on the basis of higher IL-2 secretion and lower EA oocyst production.     

Cell cycle analysis of bovine cultured somatic cells by flow cytometry

This study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells (fetal fibroblasts, adult skin and muscle cells, and cumulus cells) after culture under a variety of conditions; 1) growth to 60-70% confluency (cycling), 2) serum starvation, 3) culture to confluency. Cell -cycle phases were determined by flow cytometry with propidium iodide staining enabling the calculation of percentages of cells in G0 /G1, S and G2 /M. The majority was in G0/G1 regardless of cell type and treatment. Serum-starved or confluent cultures contained higher percentages of cells in G0/G1 (89.5-95.4%; P < 0.05). Percentages of cells in G0/G1 increased as cell size decreased regardless of the cell type and treatment. In the serum-starved and confluent cultures, about 98% of small cells were in G0/G1 . Serum-starved cultures contained higher percentages of small cells (38.5-66.9%) than cycling and confluent cultures regardless of cell type (P < 0.05) . After trypsinization of fetal fibroblasts and adult skin cells that were serum-starved and cultured to confluency, the percentages of cells in G0/G1 increased (P < 0.05) on incubation for 1.5 (95.7-99.5%) or 3 hr (95.9-98.6%). These results verify that serum starvation and culture to confluency are efficient means of synchronizing bovine somatic cells in G0/G1, and indicate that a more efficient synchronization of the cells in G0/G1 can be established by incubation for a limited time period after trypsinization of serum-starved or confluent cells.